Abstract:To clarify the flavonoid glycosyltransferase gene（RsUFGTs） involved in the biosynthesis of anthocyanin in red radish,three candidate RsUFGTs genes were screened from the radish genome,and the open reading frames (ORFs) of three RsUFGTs were cloned and sequenced and expressed. Results showed that the ORF length of RsUFGT1 and RsUFGT2 was 1 383 bp and 1 374 bp,encoding 460 and 457 amino acids,respectively. However,RsUFGT3 was mutated with early termination. Protein sequence alignment and phylogenetic tree analyses indicated that RsUFGT1 and RsUFGT2 both contain PSPG conserved domain and belong to the same UFGT class. The results of realtime quantitative PCR analysis showed that the expression of RsUFGT1 in ‘Hongxin No.1’ was relatively high and no expression was detected in skin of ‘Mantanghong’ and in skin and flesh of ‘Chunbulao’. The expression of RsUFGT1 was positively correlated with anthocyanins content in tissues of different radish cultivars. However,no obvious rules of RsUFGT2 expression were detected. It is indicated that RsUFGT1 may be one of the important structural genes of anthocyanin biosynthesis in red radish.

Abstract:In order to reveal rules of proteomic changes in different leaf positions of Populus spp.(Section tacamahaca) and senescence mechanism of the leaves,proteins expression profile at three leaf positions of Populus spp. (S. tacamahaca) hybrids were studied with labelfree proteomics.The proteins of three different leaf position were detected using labelfree method.111 proteins with differential expression were found,only 55% proteins expressed significantly differently in comparison with the 5th and 25th leaf position.Functional analysis of differentially expressed proteins showed that the proteins involved in photosynthesis,growth and development were mostly downregulated at 10th and 25th leaf position,while proteins involved in stress,nutrient decomposition and transport were mostly upregulated among the leaf senescence process.However,some proteins related to transcriptional,translation and protein modification regulation had fluctuations among the leaf senescence process.Among the three leaf positions of Populus spp.(S. tacamahaca),photosynthesis and nutrient synthesis process at 10th leaf position were more active than that at 5th leaf position.The 25th leaf leaves are enhanced by the decomposition and transport of aging nutrients,and the synthesis of the material is weakened.The decomposition and transport of senescence nutrient at 25th leaf position were enhanced and synthesis of nutrient was decreased.Protein is a cell function performer,which shows that the maturity of young leaves to senescence is a process of orderly regulating protein expression changes.

Abstract:ERF (ethylene response factor) transcription factor plays a critical role in response to environmental stress and hormonal signal transduction in plants.To elucidate the function of ERF gene in Prunus mume,the PmERF4 gene was cloned from a Prunus mume ‘Xuemei’ by RTPCR.The fulllength cDNA sequence of PmERF4 was 842 bp with an open reading frame (ORF) of 690 bp,encoding 229 amino acids.The deduced amino acid sequence alignment of PmERF4 contained a conserved AP2/ERF domain.The result of phylogenetic analysis showed the PmERF4 was highly homologous to other proteins in Prunus genus.qRTPCR was conducted to examine the expression patterns of PmERF4 under abiotic stress and hormone treatment.The results showed that PmERF4 was induced by cold and H2O2 stress.The induction levels of the gene were different between cold and H2O2 stress.A continuous and strong response of PmERF4 was triggered by cold.PmERF4 was insensitive to mannitol,MeJA and SA treatment and its expression was inhibited by NaCl and ABA.It is indicated that PmERF4 from Prunus mume may be involved in response to cold and H2O2 stress.

Abstract:Based on the transcriptome data of Dryopteris fragrans,the gene sequence of MYC2 transcription factor MYC2 was cloned and named as DfMYC2.The gene accession number was MK193871.Bioinformatics analysis of the protein sequence,fluorescence quantitative analysis of different parts and expression analysis of 0.4 mmol/L methyl jasmonate (MeJA) were performed.The results showed that the DfMYC2 gene was 3 130 bp in full length and the open reading frame (ORF) was 2 496 bp in length,encoding 831 amino acids with a relative molecular weight of 261 041.86.The leaves of D. fragrans were sprayed with 0.4 mmol/L MeJA solution,and samples were taken at 0 h,2 h,4 h,6 h,8 h,12 h,24 h and 36 h after treatment.The results of realtime quantitative PCR showed that DfMYC2 had the highest expression at 4 h after treatment.It will provide the foundation for further studying the biological function of DfMYC2 gene in D. fragrans

Abstract:In this study,insertional mutagenesis by Agrobacterium tumefaciens mediated transformation (ATMT) was used to build a T-DNA insertion library of Colletotrichum higginsianum containing a collection of 2 000 insertion mutants. From the library,development and pathogenicitydeficient mutants were screened and T-DNA insertion copies and sites were analyzed. As a result,we isolated 2 growth-deficient mutants Ch-1-E393 and Ch-1-C135 with significant reduced growth on PDA medium,12 mutants with abnormal colonies and one pathogenicitydeficient mutant Ch-1-G090. Pathogenicity assays showed that after inoculation of the pathogenic defective mutant Ch-1-G090 in Arabidopsis,the incidence of leaf diseases was significantly reduced. Under microscope,few primary hyphae were found in the leaves and no secondary hyphae was observed in Ch-1-G090. Southern blot analysis indicated that the mutant Ch-1-G090 harbored two T-DNA insertions. Border flanking sequences of T-DNAs from these mutants were recovered by InversePCR. Sequence analyses revealed that the two T-DNA insertion sites in the mutant Ch-1-G090 were coding genes for a hypothetical protein (Ch063_10682) and an RNA processing enzyme (CH063_10671).

Abstract:In this study,the transcriptome of winged and wingless Lipaphis erysimi was obtained with Illumina sequencing platform. A total of 107 992 806 cleaning reads were obtained from the mixed samples,and were assembled into 69 588 unigenes with the average length of 716 bp. A total of 44 620 184 and 47 113 716 Cleaning Reads were obtained from the winged and wingless aphid samples,respectively. Analysis of gene expression between the winged and wingless samples showed 104 significantly differentially expressed genes,of which 74 genes were upregulated and 30 genes were downregulated. Subsequently,GO annotation and KEGG pathway enrichment analysis showed that among those differentially expressed genes,Hippo signaling pathway,Thyroid hormone signaling pathway and Oxytocin signaling pathway were strongly related to the wing dimorphism and its related reproduction.

Abstract:In order to analyze the function of CreA,a carbon metabolism regulator of Penicillium italicum,which is a postharvest pathogen of citrus fruits，the PicreA gene was successfully cloned.The gene has 1 236 bp cDNA length without intron,which encodes 411 amino acids and contains two typicalzinc finger domains of transcription factors.Phylogenetic tree analysis showed that the PiCreA protein had the highest homology with Penicillium expansum.Transcriptional analysis of PicreA revealed a complex express profile in response to different carbon sources.For monosaccharide,higher expression was induced in a short period of time.With the prolongation of time,in the case of repressing carbon source such as glucose,PicreA expression showed a gradual decline and then increased.It was presumed that selfinhibition of PicreA occurred.In term of the depressing carbon source like xylose,PicreA expression level lightly decreased and gradually remained fairly high.For nonmonosaccharide carbon source such as ethanol,high level of PicreA expression was maintained.The PicreA gene is stably expressed throughout the infection process of Penicillium italicum on citrus fruits.

Abstract:The physical and chemical indicators and heavy metal content in the soil around the enterprises in key areas of Hubei Province were monitored. The monitoring data were used to establish a 3layer BP neural network model with 13 inputs,1 hidden layer and 6 outputs.The content of Mn,Co,V,Ag,Tl,Sb in the monitoring area were predicted. The Nemerow index based on the monitoring and prediction results of heavy metals was used to evaluate the pollution of the area studied. The results showed that there were different levels of exceeding standard of heavy metals in the area studied. The maximum overstandard range was 1.8156.1 times. The relative error between the prediction results of six heavy metals including Mn,Co,V,Ag,Tl and Sb and the actually tested results was ranged from 0.3% to 19.9%. Mn,V,Ag,Tl,and Sb were significantly correlated with the confidence of 99% (P＜0.01,n=11).Co was significantly correlated with confidence of 95% (P＜0.05,n=11). The BP neural network prediction model constructed had good accuracy. Based on the BP neural network model,the Nemerow pollution index not exceeding the warning limit took over a proportion of 77.3%,with the proportion of light pollution of 17.4% and the ratio of moderate to severe pollution of 4.0% each.

Abstract:The candidate genes of degradating imazethapyr were screened with integration analyses of transcriptomics and proteomics data to explore the molecular mechanism of degradating imazethapyr in Trichoderma brevicompactum. The results showed that when the basal medium with 500 mg/kg mazethapyr was used as the sole carbon source to cultivate T. brevicompactum for seven days, the highest degradation efficiency was obtained. There were 1 329 genes with significant transcriptomic differences, of which 703 were upregulated and 626 were downregulated. 21 883 peptides and 4 003 proteins were identified by proteome analyses performed with ITRAQ. The results of association analyses showed that 24 genes with differential expression and protein abundance were correspondingly changed, of which 14 were upregulated and 10 were downregulated. The association coefficient between quantitative protein and gene expression was -0.047 2. The association coefficient between significant difference protein and genes with significantly differential expression was -0.142 0, with the similar trend of expression change. The association coefficient between protein and gene expression was -0.741 3.The association coefficient between protein and genes with the opposite trend of expression change was -0.791 6. Two of the 24 genes identified by association analysis had functional annotations. They are Trichoderma virens Gv298 glycoside hydrolase family 16 protein partial mRNA and HEX1 gene, respectively. They can be used as candidate genes of degradating imazethapyr. It is indicated that the candidate genes of degradating imazethapyr by T. brevicompactum can be effectively screened with the association analyses of transcriptome and proteome.

Abstract:To study the effects of leucine on the sperm motility of yellow catfish (Pelteobagrus fulvidraco) during early stage of autumn breeding when the quality of sperms and eggs is lower than in summer breeding,males about 100 g were divided into three groups and injected with 200 μL physiological saline (control group),100 μmol/L leucine (moderate leucine treatment group) and 1 000 μmol/L leucine (high concentration leucine treatment group),respectively.It was found that sperm motility of the moderate leucine treatment group declined more slowly detected by the computer assisted sperm analysis (CASA) system.In artificial reproduction,the fertilization rate of the moderate leucine treatment group sperm was 34.91%±4.7%,significantly higher than that of the control group (15.9%±2.2%) (P＜0.05),and the fertilization rate of the high concentration leucine treatment group sperm was 16.4%±3.4%,significantly lower than that of the moderate group (P＜0.05),but not significant different from the control group (P＞0.05).The results showed that moderate leucine could enhance the sperm motility of yellow catfish,but excessive leucine could not improve the sperm reproduction.So appropriate leucine can be added to improve the sperm reproduction of yellow catfish during the early stage of autumn breeding.

Abstract:In the present study,294 Schizothoracinae specimens of Schizopygopsis pylzovi, Gymnocypris chilianensis and Schizopygopsis kialingensis,were collected from 8 sampling sites from the Yangtze River,the Yellow River and inland rivers in Gansu Province.Mitochondrial DNA Cyt b gene was used as molecular marker to perform the phylogenetic analyses,genetic diversity and population differentiation analysis of the 3 Schizothoracinae fishes.Phylogenetic analyses of all haplotypes based on NJ and BI methods revealed that S. pylzovi was clustered as a monophyletic group.However, S. kialingensis was clustered into a clad with G. chilianensis,suggesting that the two species are under early stage of speciation.Genetic diversity analysis showed that a total of 23 haplotypes were detected in the Cyt b gene of 70 S. pylzovi,with the haplotype diversity and nucleotide diversity being 0.917 and 0.003 10,respectively.A total of 28 haplotypes were detected in the Cyt b gene of 132 G. chilianensis,with the haplotype diversity and nucleotide diversity being 0.819 and 0.009 82,respectively.A total of 13 haplotypes were detected in the Cyt b gene of 92 S. kialingensis,with the haplotype diversity and nucleotide diversity being 0.706 and 0.002 33,respectively.No haplotypes were shared between either two different species,and significant population differentiation were detected among geographic populations,showing that geographic isolation has a significant impact on these Schizothoracinae fishes,and adaptive differentiation as well.Hence,different populations should be treated as different evolutionary significant units in conservation management.

Abstract:In order to study the regulation mechanism of homeostasis in vivo,Steap4 gene,which was upregulated specifically,was obtained by screening the transcriptome data of highfatfeeding mice.Polymorphism of STEAP4 among different species was compared,and the changes of autophagy related genes at transcription and translation levels were detected 24 hours after transfection in HepG2 cells.The results showed that STEAP4 was conservative among different species,their gene polymorphism mainly changed at the protein level,and the similarity of STEAP4 between mouse and human was the highest.The results of fluorescence quantitative PCR showed that mSTEAP4 had no significant effect on autophagy genes in HepG2 cells,and the results of Western blot showed that mSTEAP4 could reduce the expression of autophagy related proteins,suggesting that mSTEAP4 inhibits the autophagy at the protein level in HepG2 cells.STEAP4 could inhibit autophagy in homeostasis,reduce material and energy metabolism,and keep cells in the resting state.

Abstract:The aim was to construct a safe and efficient naringin nutrient delivery system. Whey protein isolate （WPI） was used as emulsifier,the preparation conditions were studied for naringin monolayer nanoemulsion （NA/WPΙe） and bilayer nanoemulsion with ιcarrageenan and gum arabic. Subsequently,the stability of the NA/WPΙe and naringin bilayer nanoemulsion was investigated under different environmental factors. The results showed that the optimal preparation conditions of NA/WPΙe were:5 g/100 mL WPI concentration,10% oiltoaqueous ratio（V/V）,68.95 MPa homogenization pressure for 3 cycles. The naringin encapsulation ratio could reach 75.50% under this condition.For the bilayers nanoemulsion with 0.3 g/100 mL of gum arabic and ιcarrageenan,naringin encapsulation ratios were the highest,which were 69.80% and 64.78%,respectively. The naringin bilayer nanoemulsion’s droplets were significantly smaller than NA/WPΙe at 100℃ water bath for 10 min and different NaCl concentrations （P＜0.05）. After 35 days of storage,the growth rate of bilayer nanoemulsion’s droplets was significantly lower than that of NA/WPΙe（P＜0.05）. And the storage stability increased with increasing concentration of ιcarrageenan and gum arabic. Bilayer nanoemulsion with ιcarrageenan owned higher ionic stability and storage stability than that with gum arabic. The naringin bilayer nanoemulsion improved the stability of NA/WPΙe at its isoelectric point. In conclusion,the bilayer nanoemulsion prepared by adding gum arabic and ιcarrageenan had better physical stability,so it can provide better protection for naringin.

Abstract:In order to develop new available matrixcompatible solid phase microextraction (SPME) coating and promote the development of in vivo SPME sampling technology in agriculture,the matrix compatibility of solgel polydimethylsiloxane coating was evaluated and a simple and sensitive DISPME/gas chromatography method was established for analysis of five organophosphorus pesticides in citrus pulp.When the citrus pulp was diluted by equal amount of water,adjusted to pH 1.84,and subjected to DISPME at 50℃ for 60 min with a stirring rate of 1 200 r/min,the good linear ranges were obtained with linear regression coefficients (R2) greater than 0.999 for all test compounds in citrus matrix as well as low limits of detection (ranging from 0.002 to 0.046 μg/kg),good repeatability (0.5%15.6%) and high recoveries (71.0%125.3%).The solgel fiber could be used more than 50 times for in vivo sampling in citrus pulp.

Abstract:In this experiment,artificially wounded yam tubers were treated at 5℃,15℃,25℃ and 35℃ for 7 days,respectively.The effects of different temperature treatments on the postharvest tissue structure,weight loss and disease resistance,disease resistance indicators related with metabolism of active oxygen and phenolic substance were studied.The results showed that in the range of 535℃,the corrugated polyphenols and lignin were accumulated in the wound section with the increase of treatment temperature.The treatment group with the high temperature of 35℃ accumulated the earliest and accumulated the most.The weight loss rate and incidence of yam tuber treated with callus at 35℃ were significantly reduced.The contents of hydrogen peroxide,total phenols and flavonoids,the production rate of superoxide anion,and activities of superoxide dismutase,peroxidase,polyphenol oxidase and phenylalanine ammonia lyase of the yam tuber under 35℃ were the highest among the treatments.It is indicated that the high temperature treatment of postharvest yam at 35℃ can improve the metabolism of active oxygen and phenolic of yam tubers,effectively promote the healing and disease resistance of yam tuber postharvest damage,and reduce postharvest rot.

Abstract:To investigate the potential of near infrared reflectance spectroscopy (NIRS) and chemometrics methods to quickly predict the freshness of Megalobrama amblycephala,NIR spectra of 150 samples from different seasons,different origins,different specifications and different storage durations were recorded in the range of 1 0001 799 nm. The spectra were preprocessed and then calculated using CARS for wavelength variable selection for establishing quantitative models to predict pH,TVBN,TBA and K values with partial least squares regression (PLSR) method. The results showed that correlation coefficients of the models were 0.961,0.881,0.955 and 0.946,and root mean square errors of cross validation (RMSECV) were 0.049,1.659,0.047 and 2.558,respectively. The models have good prediction ability,which will provide an effective method for predicting the freshness of freshwater fish quickly and nondestructively.

Abstract:A knapsack electric trimmer was designed to perform dynamic and kinematic analysis of the sawing process and to build a functional model based on power and torque. Taking the trimming power,torque and cutting quality as indicators,the main influencing factors were selected by theoretical analysis,and the mixed orthogonal test was carried out. The optimal combination parameters were predicted by range analysis and variance analysis. The results showed that the optimal number of cutter teeth,the diameter,and the speed was 40,180 mm,and 2 000 r/min,respectively. The test results of verification test of citrus branches with a diameter of 30 mm using an optimized pruning machine showed that the average incision quality and the torque was reduced by 2.8% and 0.3%,the power was increased by 2.6%,and the variation was small. The rationality and effectiveness of the bench test was verified. This combination of parameters can effectively reduce the trimming power and meet the trimming requirements.

Abstract:A special grassfeeding guide device for the disc lawn mower was designed for the coarse and difficulttocut weeds that are commonly grown in the orchard. Combined with CREO and ADAMS,the rigidflexible coupling model of grassfeeding guides and weeds was established,and the movement laws of weeds in the case of grassfeeding guide device with different angles and advance speeds were analyzed. According to the relationship between the displacement of the weed tip point in the X,Y and Z directions of the space coordinate system,three important indicators including the offset angle of the weed,the theoretical maximum cutting width of the mower and the cutting offset were calculated to characterize the working effect of the grassfeeding guide device. Using DesignExpert 10.0 software,a comprehensive test was designed,and the response surface effects of the three indicators were analyzed,and the experiments were verified and compared. The results showed that the optimal angle of the grassfeeding port was 80°,with the machine forward speed of 1 330 mm/s. At this time,the grassfeeding guide device worked well with flat cutting and the maximum offset of 27.02 mm.

Abstract:Crucian,grass carp,bream and yellow catfish were used to study the stress distribution of carborne live fish in the process of linear acceleration and deceleration.Using the least squares method and Gaussian function as the fitting function,the force value curves of the braking phase when traveling in a straight line of 40,60,80 km/h were respectively fitted,and the particle swarm algorithm was used to optimize the curve after fitting.The final curve had a coefficient of determination between 0.902 and 0.986,and the standard error was between 0.058 and 0.145.The fitting effect was good.The results of analyzing effects of different types of fish and vehicle speed on the fitting curve showed that the greater the speed in straightline driving,the larger the proportional coefficient and peak width parameter of the braking curve of the braking phase.Under the same situation,the squid was most affected by the braking (38.02 N) and the carp was the smallest (15.69 N).The speed of the vehicle and the type of fish in the straight line had no obvious influence on the position parameters,and the speed of the vehicle during straight running had no significant influence on the index of the fitted curve.The results of analyzing the force of the pressure test board in the braking phase showed that the measured value of the pressure in the braking phase was smaller than the theoretical value,and the relative error was less than 15.990%.

Abstract:CARMA3 (caspase recruitment domain and membraneassociated guanylate kinaselike domain protein 3) belongs to CARMA family and is a novel scaffold protein.By regulating the activation of NF-κB signaling pathway,CARMA3 controls cell cycle,proliferation and signal transduction,and consequently influences progression of cancers and cellular responses.Here,we comprehensively reviewed NF-κB signaling pathway activation process,including CARMA3mediated NF-κB activation triggered by ligands of GPCRs (G proteincoupled receptors),DNA damage or infection,influence of CARMA3mediated NF-κB activation on VEGF (vascular endothelial growth factor) activity,negative regulation of A20 on CARMA3mediated NF-κB activation,and the link between CARMA3 and cancers,aiming to provide reference for the study of cell cycle progression,proliferation and cellular responses based on NFκB signaling pathway.